Protein kinase C potentiates transmitter release from the chick ciliary presynaptic terminal by increasing the exocytotic fusion probability.

1. The giant presynaptic terminal of chick ciliary ganglion was used to examine how protein kinase C (PKC) modulates neurotransmitter release. Cholinergic excitatory postsynaptic currents (EPSCs) were recorded under whole-cell voltage clamp. 2. Although the EPSC was potentiated by phorbol ester (phorbol 12-myristate 13-acetate, PMA; 0.1 microM) in a sustained manner, the nicotine-induced current was unaffected. PMA increased the quantal content to 2.4 +/- 0.4 (n = 9) of control without changing the quantal size. 3. The inactive isoform of PMA, 4alpha-PMA, showed no significant effect on EPSCs. The PMA-induced potentiation was antagonized by two PKC inhibitors with different modes of action, sphingosine (20 microM) and bisindolylmaleimide I (10 microM). 4. When stimulated by twin pulses of short interval, the second EPSC was on average larger than the first EPSC (paired-pulse facilitation; PPF). PMA significantly decreased the PPF ratio with a time course similar to that of the potentiation of the first EPSC. 5. PMA did not affect resting [Ca2+]i or the action potential-induced [Ca2+]i increment in the giant presynaptic terminals. 6. The effect of PMA was less at 10 mM [Ca2+]o than at 1 mM [Ca2+]o. 7. When a train of action potentials was generated with a short interval, the EPSC was eventually depressed and reached a steady-state level. The recovery process followed a simple exponential relation with a rate constant of 0.132 +/- 0.029 s-1. PMA did not affect the recovery rate constant of EPSCs from tetanic depression. In addition, PMA did not affect the steady-state EPSC which should be proportional to the refilling rate of the readily releasable pool of vesicles. 8. These results conflict with the hypothesis that PKC upregulates the size of the readily releasable pool or the number of release sites. PKC appears to upregulate the Ca2+ sensitivity of the process that controls the exocytotic fusion probability.